LOCUS       sequence-205748-        5165 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter gene expression in S. rosetta.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5165)
  AUTHORS   Booth DS, Szmidt-Middleton H, King N
  TITLE     Transfection of choanoflagellates illuminates their cell biology and
            the ancestry of animal septins.
  JOURNAL   Mol Biol Cell. 2018 Dec 1;29(25):3026-3038. doi: 
            10.1091/mbc.E18-08-0514. Epub 2018 Oct 3.
  PUBMED    30281390
REFERENCE   2  (bases 1 to 5165)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1091/mbc.E18-08-0514"; journalName: "Mol Biol Cell"; date: 
            "2018-12-1- 1"; volume: "29"; issue: "25"; pages: "3026-3038"
COMMENT     Sequence Label: NK610
FEATURES             Location/Qualifiers
     source          1..5165
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(201..269,270..1061)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(419..438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1721..1740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1974..1991
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2144..2149,2150..2167,2168..2174)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2187..2209
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2206..2222
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4771..4788)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4771..4787)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4780..4802)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4996..5015)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5115..5137
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgccaagctt gtaagtagtg gtagtcgtcg aagacgtggg gtgcggggat
     2281 gtgcttgact tggtgggaga ggctggtgga agtagctgat gaaaatgaac tccaccacac
     2341 gccaactgcc acttgaacca cggaaggcag gcagcagtgc tgctgtgtgc gcttttcggc
     2401 gcggcgcgca gccacaaagc aacggcaatt gtggcgggtg attcgttcga acgctgcttt
     2461 tcataatttt cgccaatgtc tgaatcaatg tctttttcaa gggtgcagaa taacgaacat
     2521 tattttttat tgtgtggatt cagcgccaat tgcgatttgg agatgcgcta agcgcacaat
     2581 caccaccaaa cacgacattc cccgctttcc cgccaaactg attctcgcgc gaaacctgag
     2641 atgtgcaccg cgccgttaca agtgcgacca cgcacacctc ttcttgtcat ttatcatccg
     2701 catctccaag caagcaagca agcaaaccag gaccccctcc ctcgctcgtc acctcaccac
     2761 ctcccaaact taaacttcac aaccagcaca atggcccgca cgaagcagac cgcccgcaag
     2821 accaccggtg gcaaggcccc ccgcaagcag ctcgccacca aggctgcccg caagtccgcc
     2881 ccctccactg gcggcgtgaa gaagccccat cgcttccgcc ccggtaccgt cgctctgcgt
     2941 gagatccgcc gctaccagaa gtccaccgag ctgctgatcc gcaagctccc cttccagcgc
     3001 ctcgtccgcg agattgccca ggacttcaag accgacctcc gcttccagtc caccgccgtc
     3061 tctgccctgc aggaggccgc cgaggcttac ctcgtcaacc tctttgagga caccaacctc
     3121 tgcgccatcc acgcaaagcg cgtcaccatc atgcccaagg acatccagct ggcccgccgc
     3181 atccgtggcg agcgctctgg cggtagcggt ggctccatgg tctccaaggg cgaggaggac
     3241 aacatggcta tcatcaagga gttcatgcgc ttcaaggtcc acatggaggg ctccgtcaac
     3301 ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc
     3361 gccaagctca aggtgacgaa gggcggcccc ctccccttcg cctgggacat cctctccccc
     3421 cagttcatgt acggctccaa ggcttacgtc aagcaccccg ccgacatccc cgactacctc
     3481 aagctctcct tccccgaggg cttcaagtgg gagcgcgtca tgaacttcga ggacggcggc
     3541 gtcgtcaccg tcacccagga ctcctctctt caggacggcg agttcatcta caaggtcaag
     3601 ctgcgcggca ccaacttccc ctccgacggc cccgtcatgc agaagaagac gatgggttgg
     3661 gaggcttcct ccgagcgcat gtaccccgag gacggcgccc tcaagggcga gatcaagcag
     3721 cgcctcaagc tcaaggacgg cggccactac gacgccgagg tcaagaccac ctacaaggcc
     3781 aagaagcccg tccagctccc cggcgcctac aacgtcaaca tcaagctcga catcacctcc
     3841 cacaacgagg actacaccat cgtcgagcag tacgagcgtg ccgagggccg ccacagcacg
     3901 ggcggtatgg acgagcttta caagtcttaa gcgtctcacc ttctcaacat ggctcgctcg
     3961 ctcgatgtcg ttctctgaca ccttgcttgg cttgtcgtca tgaggttgct ttgtcatcac
     4021 caacgctggt tgcattttga tctgtgacac cttgaatcat cctccctccc ctcccttgtt
     4081 cttaattacc cacttggccc ttgctgtctc tcttggtctg tgccacgggt tgtcgcctcc
     4141 gcgggctcaa attgtgctca cacgtctttt atccatgctc gtgttcctga gcacttgacg
     4201 ctggcggtgt ctttgtgcgc tcgtgtgcat gggaacctct ctccctccct ctccctgtgt
     4261 gtaatggcgt tggcttaccg tggtatcgat ttgcctctgc gtaatgacac gccgcaacgg
     4321 cgtttgggga gtttccccgg cttgtcctgc ctcgcaaccc ttctcctttg gttcgttgtt
     4381 gcctctcaat acaatgatca aaccgaacat taatgtgagg accatgatgg catcctgctt
     4441 tattcaagca agtggaatga atgacacaga aacagattga tcgtcattgt gatgccgtgc
     4501 gtgtatttgt ggcggagagg aacaaaagtc acaggtggct acatttacac aacttcttgt
     4561 caatgcaaac tcagtgagtc attgggtatc atgcgtggtg aagaatgagg gagtgctgcc
     4621 agcatctgaa cgagcaccat cgtgccaggg cgcggctctt ggcaaactcg cgtgcgtcca
     4681 tctcttcgtg atcatcgtca tcatcgcgac gctgtgtcac atgtgggaca atgaatgtct
     4741 agaggatccc cgggtaccga gctcgaattc actggccgtc gttttacaac gtcgtgactg
     4801 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     4861 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     4921 cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     4981 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     5041 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     5101 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     5161 cgcga
//
